Investigation of nucleotide binding sites on chloroplast coupling factor 1 with 3'O-(4-benzoyl)benzoyl adenosine 5'-triphosphate

The subunit locations of each of the three nucleotide binding sites of soluble chloroplast coupling factor 1 have been studied with the photoaffinity label 3'-O-(4-benzoyl)benzoyl-ATP. This derivative is an effective inhibitor of ATPase activity. Photolysis of the radioactive label when bound t...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1985-04, Vol.82 (7), p.1950-1953
Main Authors: Kambouris, N G, Hammes, G G
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - analogs & derivatives
Adenosine Triphosphate - metabolism
Affinity Labels - metabolism
Binding Sites
Chloroplasts - metabolism
Nucleotides - metabolism
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Summary:The subunit locations of each of the three nucleotide binding sites of soluble chloroplast coupling factor 1 have been studied with the photoaffinity label 3'-O-(4-benzoyl)benzoyl-ATP. This derivative is an effective inhibitor of ATPase activity. Photolysis of the radioactive label when bound to each of the three nucleotide sites on the coupling factor has been examined. For the nucleotide site that normally binds ADP very tightly, NaDodSO4/polyacrylamide gel electrophoresis after photolysis indicates that primarily the beta polypeptide chain is appreciably labeled (86%), although some labeling of the alpha polypeptide chain is found (14%). For the site that binds MgATP tightly, 97% of the radioactivity is found on the beta polypeptide chain. The alpha and beta polypeptide chains are labeled in approximately equal amounts when photolysis is carried out with the nucleotide analog bound to the third site.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.82.7.1950